RNA compatible with isopentane?

Henk Veldman H.Veldman at lab.azu.nl
Thu Apr 6 02:39:42 EST 2000


soenke behrends schreef:

> Dear netters,
>
> we usually freeze our tissue
> samples in liquid nitrogen.
> From a cooperation we now get
> samples that were frozen in
> isopentane immediately after
> preparation and were then
> stored at -80C. Is it conceivable
> that the Isopentane does harm to
> RNA or protein if we want to do
> Westerns from the tissue or will
> it just work allright?
>
> Thanks for your help
> Soenke Behrends

Freezing of tissue with the aid of isopentane is much quicker than by
just dumping it in liquid nitrogen. It is done to avoid the formation of
larger ice crystals  in order to produce a better morphology in frozen
sections, especially for muscle.  Isopentane is a volatile hydrocarbon,
it is not water misceable and not reactive towards tissue (it is,
however, highly flammable).
We allways freeze our tissue samples with the use of isopentane and
never have met any problems when they later are to be used for Western
blotting,  in situ hybridisation or DNA-analysis.
So I think everything will work out fine with your samples.

Greetings, Henk




--

H. Veldman
Laboratory for Experimental Neurology (NMZ)
Room G 02.320
University Medical Center Utrecht (AZU)
P.O. Box 85500
3508 GA  UTRECHT
The Netherlands
Tel: +31 30 2507565
Fax: +31 30 2542100
E-mail: H.Veldman at lab.azu.nl






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